Xist RNA binds select autosomal genes and depends on Repeat B to regulate their expression
Xist, a pivotal player in X chromosome inactivation (XCI), has long been perceived as a cis-acting long noncoding RNA that binds exclusively to the inactive X chromosome (Xi). However, Xist’s ability to diffuse under select circumstances has also been documented, leading us to suspect that Xist RNA...
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eLife Sciences Publications Ltd
2025-06-01
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| Online Access: | https://elifesciences.org/articles/101197 |
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| author | Shengze Yao Yesu Jeon Barry Kesner Jeannie T Lee |
| author_facet | Shengze Yao Yesu Jeon Barry Kesner Jeannie T Lee |
| author_sort | Shengze Yao |
| collection | DOAJ |
| description | Xist, a pivotal player in X chromosome inactivation (XCI), has long been perceived as a cis-acting long noncoding RNA that binds exclusively to the inactive X chromosome (Xi). However, Xist’s ability to diffuse under select circumstances has also been documented, leading us to suspect that Xist RNA may have targets and functions beyond the Xi. Here, using female mouse embryonic stem cells (ES) and mouse embryonic fibroblasts (MEF) as models, we demonstrate that Xist RNA indeed can localize beyond the Xi. However, its binding is limited to ~100 genes in cells undergoing XCI (ES cells) and in post-XCI cells (MEFs). The target genes are diverse in function but are unified by their active chromatin status. Xist binds discretely to promoters of target genes in neighborhoods relatively depleted for Polycomb marks, contrasting with the broad, Polycomb-enriched domains reported for human XIST RNA. We find that Xist binding is associated with down-modulation of autosomal gene expression. However, unlike on the Xi, Xist binding does not lead to full silencing and also does not spread beyond the target gene. Over-expressing Xist in transgenic ES cells similarly leads to autosomal gene suppression, while deleting Xist’s Repeat B motif reduces autosomal binding and perturbs autosomal down-regulation. Furthermore, treating female ES cells with the Xist inhibitor, X1, leads to loss of autosomal suppression. Altogether, our findings reveal that Xist targets ~100 genes beyond the Xi, identify Repeat B as a crucial domain for its in-trans function in mice, and indicate that autosomal targeting can be disrupted by a small molecule inhibitor. |
| format | Article |
| id | doaj-art-0a18a5d9d75846efa7682f2bd5f14338 |
| institution | Kabale University |
| issn | 2050-084X |
| language | English |
| publishDate | 2025-06-01 |
| publisher | eLife Sciences Publications Ltd |
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| spelling | doaj-art-0a18a5d9d75846efa7682f2bd5f143382025-08-20T03:45:56ZengeLife Sciences Publications LtdeLife2050-084X2025-06-011310.7554/eLife.101197Xist RNA binds select autosomal genes and depends on Repeat B to regulate their expressionShengze Yao0https://orcid.org/0000-0003-4195-8402Yesu Jeon1Barry Kesner2Jeannie T Lee3https://orcid.org/0000-0001-7786-8850Department of Molecular Biology, Massachusetts General Hospital, Boston, United States; Department of Genetics, The Blavatnik Institute, Harvard Medical School, Boston, United StatesDepartment of Molecular Biology, Massachusetts General Hospital, Boston, United States; Department of Genetics, The Blavatnik Institute, Harvard Medical School, Boston, United StatesDepartment of Molecular Biology, Massachusetts General Hospital, Boston, United States; Department of Genetics, The Blavatnik Institute, Harvard Medical School, Boston, United StatesDepartment of Molecular Biology, Massachusetts General Hospital, Boston, United States; Department of Genetics, The Blavatnik Institute, Harvard Medical School, Boston, United StatesXist, a pivotal player in X chromosome inactivation (XCI), has long been perceived as a cis-acting long noncoding RNA that binds exclusively to the inactive X chromosome (Xi). However, Xist’s ability to diffuse under select circumstances has also been documented, leading us to suspect that Xist RNA may have targets and functions beyond the Xi. Here, using female mouse embryonic stem cells (ES) and mouse embryonic fibroblasts (MEF) as models, we demonstrate that Xist RNA indeed can localize beyond the Xi. However, its binding is limited to ~100 genes in cells undergoing XCI (ES cells) and in post-XCI cells (MEFs). The target genes are diverse in function but are unified by their active chromatin status. Xist binds discretely to promoters of target genes in neighborhoods relatively depleted for Polycomb marks, contrasting with the broad, Polycomb-enriched domains reported for human XIST RNA. We find that Xist binding is associated with down-modulation of autosomal gene expression. However, unlike on the Xi, Xist binding does not lead to full silencing and also does not spread beyond the target gene. Over-expressing Xist in transgenic ES cells similarly leads to autosomal gene suppression, while deleting Xist’s Repeat B motif reduces autosomal binding and perturbs autosomal down-regulation. Furthermore, treating female ES cells with the Xist inhibitor, X1, leads to loss of autosomal suppression. Altogether, our findings reveal that Xist targets ~100 genes beyond the Xi, identify Repeat B as a crucial domain for its in-trans function in mice, and indicate that autosomal targeting can be disrupted by a small molecule inhibitor.https://elifesciences.org/articles/101197Xist RNARepeat BX1 inhibitorautosomal targets |
| spellingShingle | Shengze Yao Yesu Jeon Barry Kesner Jeannie T Lee Xist RNA binds select autosomal genes and depends on Repeat B to regulate their expression eLife Xist RNA Repeat B X1 inhibitor autosomal targets |
| title | Xist RNA binds select autosomal genes and depends on Repeat B to regulate their expression |
| title_full | Xist RNA binds select autosomal genes and depends on Repeat B to regulate their expression |
| title_fullStr | Xist RNA binds select autosomal genes and depends on Repeat B to regulate their expression |
| title_full_unstemmed | Xist RNA binds select autosomal genes and depends on Repeat B to regulate their expression |
| title_short | Xist RNA binds select autosomal genes and depends on Repeat B to regulate their expression |
| title_sort | xist rna binds select autosomal genes and depends on repeat b to regulate their expression |
| topic | Xist RNA Repeat B X1 inhibitor autosomal targets |
| url | https://elifesciences.org/articles/101197 |
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